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succinus seem to be frequent causes of IMI as well as contaminants. If a species-differentiation is available after cultivating NAS, the findings should be interpreted in consideration of the observations made in this study, whether it is more likely a question of a contaminant or a cause of intramammary infection. The bacteria shedding intensity of the NAS species with a more substantially adverse effect on udder health seems to be higher than that of the less important NAS pathogens. A total of 96 Trueperella pyogenes isolates, an opportunistic pathogen of food-producing ruminants, obtained from cattle (n = 34), sheep (n = 35) and goats (n = 27), and identified by Real Time PCR (qPCR), were analysed to determine the susceptibility to 12 antimicrobials commonly used in livestock, using a broth microdilution. The Minimal Inhibitory Concentration (MIC) distribution was unimodal for half of the antimicrobials tested with the exception of apramycin, gentamicin, streptomycin, oxytetracycline, tylosin, and erythromycin all of which showed bimodal MIC distributions. Low MIC90 values for penicillin, amoxicillin, ceftiofur, enrofloxacin, and gentamicin ( less then 1 μg/ml) were obtained, suggesting that these antimicrobials would be the most effective first line empiric treatment for T. pyogenes infections in livestock. Furthermore, according to the specific T. pyogenes breakpoints for penicillin, sulfamethoxazole/trimethoprim and erythromycin, 93.7 % of isolates were susceptible to penicillin and 77.2 % to erythromycin, whereas 92.7 % were non-susceptible to sulfamethoxazole/trimethoprim. Significant differences were observed in the MIC distribution of almost all antimicrobials, except enrofloxacin, tylosin and erythromycin against cattle, sheep or goat isolates, although all antimicrobials showed similar MIC90 values, except apramycin and oxytetracycline that showed higher values when tested against cattle isolates. These data provide interesting information on the antimicrobials of choice for the treatment of infections caused by T. pyogenes in ruminants. A number of veterinary clinical pathology laboratories in New Zealand have been reporting emergence of increased minimum in inhibitory concentrations for β-lactams in the common clinical bovine mastitis pathogen Streptococcus uberis. The objective of this study was to determine the genetic basis of this increase in MIC for β-lactams amongst S. uberis. Illumina sequencing and determination of oxacillin MIC was performed on 265 clinical isolates. Published sequences of the five penicillin binding proteins pbp1a, pbp1b, pbp2a, pbp2b, and pbp2x were used to identify, extract and align these sequences from the study isolates. Amino acid substitutions resulting from single nucleotide polymorphisms (SNP) within these genes were analysed for associations with elevated (≥ 0.5 mg/L) oxacillin MIC together with a genome wide association study. The population structure of the study isolates was approximated using a phylogenetic tree generated from an alignment of the core genome. A total of 53 % of isolates had MIC ≥ 0.5erapy for clinical mastitis. Pigeon circovirus (PiCV) is the most diagnosed virus in pigeons (Columba livia) and have been studied and reported globally. PiCV infections can lead to immunosuppression and pigeons infected with PiCV can result to lymphocyte apoptosis and atrophy of immune organs. Young pigeon disease syndrome (YPDS) is a complex disease and believed that PiCV could be one of the agents leading to this syndrome. An effective treatment regimen is needed to control the spread of PiCV in pigeons. selleck inhibitor In this study pigeon interferon alpha (PiIFN-α) was cloned and expressed and its antiviral effects were tested against fowl adenovirus type 4 (FAdV-4) in vitro and PiCV in vivo. No detectable levels of FAdV-4 viral genome in LMH cells stimulated with 300 μg/mL PiIFN-α were found. Additionally, PiIFN-α was stable at different temperature and pH for 4 h, and no reduction in antiviral activity was observed in untreated and treated cells. In pigeons naturally and experimentally infected by PiCV, no detectable levels of PiCV virus titers were found after treatment with PiIFN-α. Cytokine and ISG expression levels in liver and spleen samples were detected and IFN-γ and Mx1 genes were dominantly up-regulated following PiIFN-α treatment (p less then 0.05). This study demonstrated that PiCV can be inhibited by administration of PiIFN-α and PiFN-α can be used as a therapeutic approach to prevent the spread of PiCV in pigeons. Recombinant Muscovy duck parvovirus (rMDPV) has been recently identified as a novel pathogen circulating in Chinese Muscovy duck flocks in the past two decades. Different from classical MDPV, rMDPV infection can form embolism in the intestinal tract of deceased Muscovy ducklings. However, whether rMDPV acts as the sole causative agent involved in the formation of the characteristic embolism in Muscovy ducklings remains unclear. In this study, an infectious plasmid clone pZW containing the complete genome of strain ZW, a previously characterized rMDPV isolate, was constructed, and a single nucleotide mutation was then introduced in the VP1 gene within pZW as the genetic marker. Transfection of pZW in 11-day-old embryonated Muscovy duck eggs via the chorioallantoic membrane route resulted in the rescue of the infectious virus. The rescued virus exhibited similar biological characteristics to its parental strain ZW, as evaluated by the median embryo lethal dose and the replication kinetics in embryonated Muscovy duck eggs. Muscovy duckling infection tests showed that the rescued virus and parental strain can kill all Muscovy ducklings within 7 days post-infection. Postmortem examination revealed that embolism can be observed in the intestinal tracts of deceased ducklings in the rescued and parental virus infection groups. Collectively, the present study demonstrated that sole rMDPV infection of Muscovy ducklings, without participation of other pathogens, is enough to form characteristic embolism in the intestinal tract. The CRISPR/CRISPR-associated protein 9 (Cas9) system is a powerful gene-editing tool originally discovered as an integral mediator of bacterial adaptive immunity. Recently, this technology has been explored for its potential utility in providing new and unique treatments for viral infection. Marek's disease virus (MDV) and avian leukosis virus subgroup J (ALV-J), major immunosuppressive viruses, cause significant economic losses to the chicken industry. Here, we evaluated the efficacy of using MDV as a CRISPR/Cas9-delivery system to directly target and disrupt the reverse-transcribed products of the ALV-J RNA genome during its infection cycle in vitro and in vivo. We first screened multiple potential guide RNA (gRNA) target sites in the ALV-J genome and identified several optimized targets capable of effectively disrupting the latently integrated viral genome and providing efficient defense against new infection by ALV-J in cells. The optimal single-gRNAs and Cas9-expression cassettes were inserted into the genome of an MDV vaccine strain.
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